Development of a chemically-defined minimal medium for studies on growth and protein uptake of Gemmata obscuriglobus

Abstract

We experimentally determined minimal media requirements for Gemmata obscuriglobus, a Gram-negative Planctomycete bacteria with several unusual physiological features. We find that supplementing media with the usual vitamins solution does not improve viability, but does result in an increased growth rate in liquid cultures and a larger colony size on agar plates. By systematically including individual vitamins, or omitting individual vitamins, from media we find that the addition of only two vitamins, biotin and cyanocobalamin, are sufficient to restore colony growth to comparable rates as other commonly used media. Overall, our findings define minimal media requirements for the culturing of this low-nutrient organism. One of G. obscuriglobus unusual physiological features is the ability to internalize fully-folded proteins. Using fluorescence microscopy and flow cytometery we show that this physiological behavior is dependent on media state and composition. The percentage of cells exhibiting internalization of GFP when grown on a particular, solid minimal medium is far greater than cells grown in liquid medium of similar composition or other solid media with different compositions.